Inter- and intraobserver agreement in whole-slide digital ThinPrep samples of low-grade squamous lesions of the cervix uteri with known high-risk HPV status: A multicentric international study.

BACKGROUND: High-risk human papilloma virus (HR HPV) testing and liquid-based cytology are used for primary cervical screening. Digital cytology, based on whole-slide scanned samples, is a promising technique for teaching and diagnostic purposes. The aim of our study was to evaluate the interobserver and intraobserver variation in low-grade squamous lesions, HR HPV status bias, and the use of whole-slide scanned digital cervical cytology slides. METHODS: Fifteen expert cytopathologists evaluated 71 digitalized ThinPrep slides (31 atypical squamous cells of undetermined significance [ASC-US], 21 negative for intraepithelial lesion or malignancy, and 19 low-grade squamous intraepithelial lesion cases). HR HPV data were accessible only in the second round. RESULTS: In interobserver analysis, Kendall's coefficient of concordance was 0.52 in the first round and 0.58 in the second round. Fleiss' kappa values were 0.29 in the first round and 0.31 in the second round. In the ASC-US category, Fleiss kappa increased from 0.19 to 0.22 in the second round and the increase was even higher expressed by Kendall's coefficient: from 0.42 to 0.52. In intraobserver analysis, personal scores were higher in the second round. CONCLUSIONS: The interobserver and intraobserver variability in low-grade squamous lesions was within fair agreement values in the present study, in line with previous works. The comparison of two rounds showed that expert cytopathologists are generally unbiased by the knowledge of HR HPV data, but that being informed of the HR HPV status leads to a better agreement. Stain quality and back discomfort were highlighted as factors affecting digital cytopathology use.

The newly proposed International Endocervical Adenocarcinoma Criteria and Classification and its relevance to cervical cytology screening assessed in a prospective 2-year study of 118 cases.

BACKGROUND: It is generally acknowledged that interobserver variability for the histological diagnosis of endocervical adenocarcinoma (EA) subtypes is suboptimal. The recently proposed International Endocervical Adenocarcinoma Criteria and Classification (IECC) system is based on the presence of associated human papilloma virus (HPV) infection. It recognises HPV-associated EAs and non-HPV-associated EAs. METHODS: This prospective cytology-histology and molecular genetics-based study investigated the potential effect of IECC being applied to Papanicolaou (Pap) test with regard to the diagnostic accuracy of severe glandular lesions reported at least as adenocarcinoma in situ (AIS). RESULTS: Out of 118 liquid-based cytology Pap tests with AIS+ lesion, complete information on follow-up biopsy and HPV status was available in 51 cases. AIS and EA category correlated with histologically confirmed AIS/EA in 88.5% (23/26) and 70.5% (12/17) of cases, respectively. Interestingly, 93% (40/43) of cases diagnosed as AIS/EA were HPV positive and 7% (3/43) were HPV negative (originating in the cervix, endometrium and adnexa). CONCLUSIONS: Our findings suggest that this approach could possibly divide Pap tests containing severe glandular lesion into two groups: (a) robust diagnosis of HPV-associated EA and (b) non-HPV associated glandular lesions of heterogeneous origin, requiring further clinical preoperative diagnostic workup.

ZC3H7B-BCOR high-grade endometrial stromal sarcoma may present as myoma nascens with cytoplasmic signet ring cell change.

We report on 51-year-old woman who presented with brown discharge and postcoital bleeding due to myoma nascens-like polypoid mass distending cervical canal. Histologically, the tumor consisted of high-grade spindle cell component with up to 15 mitotic figures per 10 HPF and also low-grade leiomyoma-like areas with focal myxoid change and so far undescribed cytoplasmic signet ring cell change. Immunohistochemically Desmin, actin, and h-caldesmon were negative. Conversely, BCOR positive expression was coupled with Cyclin D1 positivity and was antibody clone dependent. The molecular NGS and FISH study identified reciprocal fusion gene ZC3H7B-BCOR. In conclusion, these findings further support the idea of routine reflex molecular testing of uterine mesenchymal tumors with unusual clinical presentation or in case malignancy is suspected. Lastly, we suggest ZC3H7B-BCOR rearranged high-grade endometrial stromal sarcoma might be considered as a tumor suitable for BCL6-targeted treatment.

Suicides by ingestion of pure caffeine powder. New challenge for public health?

OBJECTIVES: Intentional or unintentional caffeine abuse due to excessive intake of energy beverages is frequent. Fatalities due to caffeine intoxication are presented rarely. Over the last years, cases of caffeine ingestion for purposes of committing suicide have been reported in scientific literature. DESIGN: The aim of this case study is to report a fatal case of a 26-year-old man who intentionally ingested pure caffeine powder. METHODS: Standard autopsy, microscopic examination and toxicological examination using screening method, TLC, GC, and GC/MS were performed. RESULTS AND CONCLUSIONS: The presence of caffeine only at the concentration of 362 ug/mL in blood and 187 ug/mL in urine was revealed. The cause of death was cardiac arrest most likely due to ventricular dysrhythmia. The concentration of caffeine determined in the femoral blood of the decedent was three times higher than the lethal concentrations reported in other published case studies. Based on the investigated circumstances, the manner of death was evaluated as a suicide.

Occurrence of periadrenal brown adipose tissue in adult Slovak population.

OBJECTIVES: The brown adipose tissue is distributed only in specific locations of the mammalian body. Evidence has been presented that the brown fat occurs in human foetuses and new-borns. At later stages of life, most brown fat cells disappear and only persist in the perirenal and periaortic adipose tissue. However, detailed data on the occurrence of brown adipose tissue in adult humans differ among existing studies. DESIGN: The aim of this study was to determine the prevalence of brown adipose tissue in adult Slovak population. SUBJECTS AND METHODS: The samples of periadrenal adipose tissue were taken from 379 consecutive autopsies of subjects aged between 26 and 94 years, fixed in formalin, processed by routine methods and stained with hematoxylin-eosin. The sections were examined by light microscopy. RESULTS AND CONCLUSIONS: Brown adipose tissue cells were found in 240 (63%) cases. The prevalence of brown adipose tissue decreases below the age of 50 and later slightly increases. No significant differences in the prevalence of brown adipose tissue between men and women were found. With the exception of Germany, this study is currently the first extensive morphological autopsy study of brown adipose tissue prevalence in adults in Central and Eastern Europe.

The detection of DNA methylation of tumour suppressor genes in cervical high-grade squamous intraepithelial lesion: A prospective cytological-histological correlation study of 70 cases.

BACKGROUND: DNA methylation has been suggested as one of the epigenetic changes promoting carcinogenesis. The aim of this study was to prospectively evaluate the methylation status of CADM 1, MAL and hsa-miR-124 genes in high-grade squamous intraepithelial lesion (HSIL) liquid-based cytology (LBC) samples with a histological correlation. METHODS: Seventy histologically confirmed cases of HSIL paired with prior screening LBC diagnosis of HSIL within a 3-month interval were selected. Histologically, the lesions were reviewed and assessed including: (a) number of blocks harbouring dysplastic squamous epithelium; (b) number of blocks containing glandular extension of dysplastic epithelium; and (c) the depth of glandular extension (which was assessed semi-quantitatively as graded 1-3). Human papillomavirus (HPV) subtyping was performed from residual LBC materials using the LINEAR ARRAY HPV Genotyping Test and in-house polymerase chain reaction targeting the HPV E1 gene. The detection of methylation silencing of tumour suppressor genes CADM1, MAL and hsa-miR-124 was performed by multiplex methylation-specific real-time polymerase chain reaction. RESULTS: A positive methylation status was detected in 41 cases (58.6%). The number of blocks with HSIL varied from one to 13. Glandular extension was seen in 44 cases with the number of blocks involved ranging from one to 10. The depth of HSIL glandular extension varied. CONCLUSION: The DNA methylation test allows HSIL lesions to be divided into two distinct groups of methylated HSIL in significantly older patients and unmethylated HSIL in younger patients. This study was not able to prove that methylation status in cervical HSIL correlates with the size of the lesion (measured by the number of blocks involved) or with HSIL propensity for endocervical glandular extension, nor with HPV type or multi-infection.

Sudden and unexpected death due to intracranial sellar extramedullary plasmacytoma.

The incidence of sudden death caused by undiagnosed primary intracranial neoplasm in young adults is extremely low. The aim of the work is to present a case report of the sudden death of a 24-year-old woman. The autopsy has revealed enlargement of sella turcica with an intrasellar tumorous mass extending into the adjacent basal parts of the brain. A tumour was diagnosed as an extramedullary plasmacytoma. The cause of death was established as a failure of the central nervous system. Toxicological analyses of biological materials were negative. According to the case history, a woman reportedly had no serious health problems. Finding of an extensive intracranial tumour in the sella turcica was random and surprising. The presented case is an extremely rare case of sudden death caused by intracranial intrasellar extramedullary plasmacytoma without previous clinical manifestation.

Long-term self-inflicted craniocerebral penetrating injury with a fatal outcome.

A case of long-term continual self-inflicted penetrating craniocerebral injury to a 59-year-old man with a fatal outcome is reported. The man suffered from paranoid psychosis and alcohol dependence. Over five months, he continually scalped the skin and subcutaneous tissue of his head with a knife until he perforated the skull and dura mater and injured his brain. He eventually sought medical advice, but died after 10 days of hospitalization despite complex conservative treatment. The immediate cause of death was a combination of central nervous system failure and heart failure. The original case presented is sporadic in the forensic literature due to atypical long-term continual self-harm to the head and brain using a sharp object that resulted in perforation of the skull and a fatal outcome. This case also highlights the importance of autopsy to determine the mechanism of injury and cause of death.

Immunohistochemical and selected genetic reflex testing of all uterine leiomyosarcomas and STUMPs for ALK gene rearrangement may provide an effective screening tool in identifying uterine ALK-rearranged mesenchymal tumors.

We examined the value of targeted molecular screening for the identification of uterine anaplastic lymphoma kinase (ALK)-rearranged mesenchymal tumors, including ALK immunohistochemistry followed by molecular genetic testing, in all uterine leiomyosarcomas and STUMPs (smooth muscle tumors of uncertain malignant potential). All leiomyosarcoma and STUMP cases diagnosed in a 10-year period (2006-2016) at Charles University Faculty of Medicine in Pilsen were retrieved and reviewed. Of 23 cases, one case (LMS [leiomyosarcoma]) was positive for ALK rearrangement, namely, PPP1CB-ALK fusion gene. No specific histologic features (i.e., lymphocytic infiltrate and stromal edema) were observed in this case. This suggests that inflammatory myofibroblastic tumor (IMT)-like histologic features may not be an initial reliable screening tool in identifying uterine IMT cases. Thus, we proposed a two-step IHC and molecular genetic testing (as a reflex test) for IMT in all uterine LMS and STUMP cases. This will enhance the proper detection of such tumors at the population level and ultimately offer patients available targeted therapies.

Accidental fatal craniocerebral injury caused by broken chain of sawing tool.

The case of an accidental fatal penetrating craniocerebral injury to a 26-year-old man at work when cutting wood is reported. There was used an angle grinder with an accessory for cutting similar to a chainsaw. The injury was caused by a fragment of the broken chain. The chain fragment broke through the facial part of the head to the left and penetrated the brain in the region of the left parietal bone of the cranial vault. The immediate cause of death was a failure of the central nervous system (brain death). The toxicological analysis of biological materials was negative. The death occurred as a result of a triple violation of safety precautions. The presented case is extremely rare in terms of fatal injuries caused by power tools for sawing having been published in the forensic literature.

[Periosteal osteosarcoma - personal experience with five cases]. / Periostální osteosarkom - osobní zkusenost s peti prípady.

The authors present five cases of periosteal osteosarcoma located in the femur (4) and tibia (1) in children and young adults (1 female and 4 males) with an age range of 9 - 23 years (mean age 15 years). Radiographs in all cases showed a broad-based soft tissue mass attached to the cortex with periosteal reaction and in two of them cortical disruption with extensive medullary involvement. Follow-ups were available in four cases (range 11 - 73 months) and revealed pelvic metastasis after 15 months with ultimately rapid dissemination and death in a 9-year-old girl and metastasis to the humerus after 13 months in a 15-year-old boy. The former tumor widely extended into the medullary cavity and an amputation was carried out, the latter had a pure juxtacortical position and an en block resection was performed; both of them were treated with chemotherapy. All the lesions displayed distinctive structural patterns combining a large island of tumorous cartilage and hypocellular, bland-looking myxoid mesenchymal stroma with abrupt transition between both components. Contrary to conventional osteosarcoma, the delicate flocculent osteoid deposits were produced by innocuous stromal cells lacking apparent atypia. They were strictly situated outside the prevailing chondroid areas and disclosed sometimes only after a meticulous search. Immunohistochemical detection of SATB2, S100protein and D2-40 assisted effectively not only in recognition of the real stromal histogenetic derivation, but also in distinction of true differentiation of a heavily mineralized extracellular matrix. Molecular analysis revealed no IDH1/2 mutation in four examined cases. Regardless of unique low-grade morphology in rare periosteal osteosarcoma, an aggressive therapeutical approach similar to conventional osteosarcoma is justified, particularly in the case of a medullary extension.